DJ-1过表达通过PTEN/Akt通路促进人胃癌MGC803细胞的增殖、迁移、侵袭与上皮间质转化  

DJ-1 over-expression promotes proliferation,migration,invasion and epithelial mesenchymal transformation of human gastric cancer MGC803 cells through PTEN/Akt pathway

作　　者：周娟[1] 夏红[1] 刘芳[1] 苏坚 苏波[3] 苏琦[1] ZHOU Juan;XIA Hong;LIU Fang;SU Jian;SU Bo;SU Qi(Institute of Cancer Research,Hunan Provincial Key Laboratory of Cancer Cell and Molecular Pathology,University of South China,Hengyang 421001,Hunan,China;Department of Pathology,the Second Affiliated Hospital,Hunan Clinical Research Center for Gastric Cancer Prevention and Treatment,University of South China,Hengyang 421001,Hunan,China;Institute of Pharmacy and Pharmacology,Key Laboratory for Pharmacoproteomics of Hunan Provincial University,University of South China,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学肿瘤研究所,湖南省肿瘤细胞与分子病理学重点实验室,湖南衡阳421001 [2]南华大学附属第二医院病理科,湖南省胃癌防治临床研究中心,湖南衡阳421001 [3]南华大学药学与药理学研究所,湖南省高校药物蛋白质组学重点实验室,湖南衡阳421001

出　　处：《中国肿瘤生物治疗杂志》2023年第7期577-585,共9页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金(No.81973532);湖南省教育厅科学研究项目(No.19C1610);南华大学科研基金项目(No.220XNK002)。

摘　　要：目的:探讨DJ-1基因过表达对人胃癌MGC803细胞增殖、迁移、侵袭与上皮间质转化(EMT)的影响及其机制。方法:利用基因转染技术构建DJ-1基因过表达MGC803细胞,实验分为MGC803、空载体和DJ-1过表达组。采用MTT、平板克隆形成、细胞划痕和Transwell实验分别检测DJ-1过表达对MGC803细胞增殖、克隆形成、迁移与侵袭的影响;qPCR和WB法检测DJ-1过表达对各组细胞DJ-1、PTEN、Akt、p-Akt、Snail、vimentin、E-cadherin、MMP-9与TIMP-3表达的影响,相差显微镜下观察MGC803细胞形态学的变化。裸鼠荷瘤实验检测DJ-1过表达对MGC803细胞移植瘤体内生长的影响。结果:成功构建DJ-1基因稳定过表达的MGC803细胞。与MGC803组和空载体组比较,DJ-1过表达组细胞的增殖能力与克隆形成数均显著增加(均P<0.05),细胞迁移距离明显增加、划痕距离明显缩短(均P<0.05),迁移与侵袭细胞数显著增多(均P<0.05),DJ-1 mRNA与蛋白表达明显上调、PTEN mRNA与蛋白表达下调(均P<0.05),Akt总蛋白各组比较无明显差异(均P>0.05),p-Akt蛋白表达明显上调(P<0.05),Snail、vimentin与MMP-9表达上调、E-cadherin与TIMP-3表达下调(均P<0.05)。相差显微镜下见长梭形细胞数目增多,圆形与椭圆形细胞减少,异型性更为明显。荷瘤裸鼠体内实验结果表明,与MGC803组相比较,DJ-1过表达组MGC803细胞移植瘤生长速度明显加快、移植瘤质量显著增加(均P<0.05)。结论:DJ-1过表达可通过PTEN/Akt通路在体内外抑制MGC803细胞的增殖、迁移、侵袭与EMT。Objective:To investigate the effects of DJ-1 gene over-expression on proliferation,migration,invasion and epithelial-mesenchymal transformation(EMT)of human gastric cancer MGC803 cells and the underlying mechanism.Methods:MGC803 cells with DJ-1 over-expression were constructed by gene transfection technology.Three groups of cells,namely MGC803 group,empty vector group,and DJ-1 over-expression group were set.The effects of DJ-1 gene over-expression on proliferation,clone formation,migration and invasion of MGC803 cells were detected by MTT,plate cloning assay,cell scratch assay and Transwell invasion assay,respectively.The effects of DJ-1 over-expression on the expression levels of DJ-1,PTEN,Akt,p-Akt,Snail,vimentin,E-cadherin,MMP-9 and TIMP-3 were detected by qPCR and Western blot.The morphological changes of MGC803 cells were observed by phase contrast microscope.The effect of DJ-1 over-expression on the growth of MGC803 cell transplanted tumor in vivo was detected in nude mice.Results:MGC803 cells with stable DJ-1 over-expression were constructed successfully.The proliferation ability and number of clones in DJ-1 over-expression group were significantly increased compared with those in MGC803 cell group and empty vector group(all P<0.05);the cell migration distance was significantly increased while the scratch distance was significantly shortened in DJ-1 over-expression group compared with those in MGC803 cell group and empty vector group(all P<0.05);and the migrated and invaded cells of DJ-1 over-expression group were significantly more than those of MGC803 group and empty vector group(all P<0.05).Moreover,the expression of DJ-1 was significantly up-regulated while the expression of PTEN was significantly down-regulated at both the mRNA and protein levels in the DJ-1 over-expression group compared with those in MGC803 group and empty vector group(both P<0.05).There was no significant difference in total Akt protein among all groups(P>0.05),but the expression of p-Akt protein in DJ-1 over-expression group was s

关 键 词：胃癌 MGC803细胞 DJ-1基因 PTEN/Akt通路 增殖 迁移 侵袭 上皮间质转化 

分 类 号：R735.2[医药卫生—肿瘤] R730.2[医药卫生—临床医学]